![DNase I, recombinant ADAMTS13, and anfibatide inhibit thrombus and prevent NET accumulation under flow in whole blood of Adamts13-/- mice. (A-C) The final surface coverage of platelets and leukocytes on a collagen surface following perfusion of Adamts13-/- murine blood samples stimulated with Stx-2 in the presence (top) or absence (bottom) of DNase I (200 IU/mL) or recombinant ADAMTS13 (+rA13) (12 μg/mL), or anfibatide (+ANF) (1.5 μg/mL), respectively, under 100 dyne/cm2. (D-F) The rate of thrombus formation over time (mean ± standard error of the mean [SEM]) following perfusion of Adams13-/- murine blood samples stimulated with Stx-2 in the presence (red) or absence (blue) of DNase I or rA13, ANF, under flow. (G-I) Representative confocal microscopic images demonstrate adhered platelets (purple), neutrophils (green), and extracellular DNA strings (elongated green) at the end of perfusion of whole blood samples from adamts13-/- mice in the presence (top) or absence (bottom) of DNase I, rA13, or ANF, following fixation and additional staining. (Gi-Ii) Enlarged (5×) areas from the bottom of panels G, H, and I, respectively, indicate the extracellular DNA strings, platelets, and neutrophils. (J-L) Number of elongated DNA strings formed at the end of perfusion of blood samples from Adamts13-/- mice in the presence or absence of DNase I, rA13, and ANF (n = 3), respectively. Data are shown as the median (bar) and IQR. Mann-Whitney test was used for the differences between the 2 groups. rA13, recombinant ADAMTS13; ANF, anfibatide; N, neutrophil; PLT, platelet. ∗∗ indicates P < .01.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodadvances/8/10/10.1182_bloodadvances.2023011617/2/blooda_adv-2023-011617-gr3.jpeg?Expires=1736058364&Signature=ImLe60xT0fAtXAbU4V7Qvq7RufB4AOgsKsFcLpTqH5kgZmYHB0axAP8DbHul9NIgJPY5wRHrA79zhl1WNvcc1wW7nlrOq2dwBMqfmscpI0ntwp70JNXnk-jalN~I6V9vluJwi4AhxgPemUGD5NL1hCklTsFunLQ-E2wV~54KEUxBpy8nAiKRgoGSnYdOqH41NSA71rjJiNfMgjEOuyxv1g5DLMd6Qh-x5pANZeNeUsO2UcwZYnkPqcUMK2oPueCwcgkmHVo2afsk9bAuWJ6aPxTVsu6oSi1vWLRXnyJaLEXK3HV5~SLXf37YqmTxLispBDcIQQmj6OZ~mcD8XMP3rQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
DNase I, recombinant ADAMTS13, and anfibatide inhibit thrombus and prevent NET accumulation under flow in whole blood of Adamts13-/- mice. (A-C) The final surface coverage of platelets and leukocytes on a collagen surface following perfusion of Adamts13-/- murine blood samples stimulated with Stx-2 in the presence (top) or absence (bottom) of DNase I (200 IU/mL) or recombinant ADAMTS13 (+rA13) (12 μg/mL), or anfibatide (+ANF) (1.5 μg/mL), respectively, under 100 dyne/cm2. (D-F) The rate of thrombus formation over time (mean ± standard error of the mean [SEM]) following perfusion of Adams13-/- murine blood samples stimulated with Stx-2 in the presence (red) or absence (blue) of DNase I or rA13, ANF, under flow. (G-I) Representative confocal microscopic images demonstrate adhered platelets (purple), neutrophils (green), and extracellular DNA strings (elongated green) at the end of perfusion of whole blood samples from adamts13-/- mice in the presence (top) or absence (bottom) of DNase I, rA13, or ANF, following fixation and additional staining. (Gi-Ii) Enlarged (5×) areas from the bottom of panels G, H, and I, respectively, indicate the extracellular DNA strings, platelets, and neutrophils. (J-L) Number of elongated DNA strings formed at the end of perfusion of blood samples from Adamts13-/- mice in the presence or absence of DNase I, rA13, and ANF (n = 3), respectively. Data are shown as the median (bar) and IQR. Mann-Whitney test was used for the differences between the 2 groups. rA13, recombinant ADAMTS13; ANF, anfibatide; N, neutrophil; PLT, platelet. ∗∗ indicates P < .01.
